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Journal of Bacteriology, May 2000, p. 2869-2878, Vol. 182, No. 10
Institute of
Microbiology1 and Protein Chemistry
Laboratory,2 Swiss Federal Institute of
Technology, ETH-Zentrum, CH-8092 Zürich, Switzerland, and
School of Biological Sciences, University of Manchester,
Manchester M13 9PT, United Kingdom3
Received 7 October 1999/Accepted 15 February 2000
Pseudomonas putida S-313 can utilize a broad range of
aromatic sulfonates as sulfur sources for growth in
sulfate-free minimal medium. The sulfonates are cleaved
monooxygenolytically to yield the corresponding phenols.
miniTn5 mutants of strain S-313 which were no longer able
to desulfurize arylsulfonates were isolated and were found to
carry transposon insertions in the ssuEADCBF operon, which
contained genes for an ATP-binding cassette-type transporter
(ssuABC), a two-component reduced flavin
mononucleotide-dependent monooxygenase (ssuED) closely
related to the Escherichia coli alkanesulfonatase, and a
protein related to clostridial molybdopterin-binding proteins
(ssuF). These mutants were also deficient in growth with a
variety of other organosulfur sources, including aromatic and aliphatic
sulfate esters, methionine, and aliphatic sulfonates other than
the natural sulfonates taurine and cysteate. This pleiotropic phenotype was complemented by the ssu operon, confirming
its key role in organosulfur metabolism in this species. Further
complementation analysis revealed that the ssuF gene
product was required for growth with all of the tested substrates
except methionine and that the oxygenase encoded by
ssuD was required for growth with sulfonates or
methionine. The flavin reductase SsuE was not required for growth with
aliphatic sulfonates or methionine but was needed for growth
with arylsulfonates, suggesting that an alternative isozyme
exists for the former compounds that is not active in transformation of
the latter substrates. Aryl sulfate ester utilization was catalyzed by
an arylsulfotransferase, and not by an arylsulfatase as in the related
species Pseudomonas aeruginosa.
0021-9193/00/$04.00+0
Copyright © 2000, American Society for Microbiology. All rights reserved.
The ssu Locus Plays a Key Role in
Organosulfur Metabolism in Pseudomonas putida
S-313
*
Corresponding author. Mailing address: School of
Biological Sciences, University of Manchester, Stopford Bldg., Oxford
Rd., Manchester M13 9PT, England. Phone: 44-161-275 3895. Fax:
44-161-275 5656. E-mail: michael.kertesz{at}man.ac.uk.
Present address: Intervet International B.V., NL-5830 AA
Boxmeer, The Netherlands.
Journal of Bacteriology, May 2000, p. 2869-2878, Vol. 182, No. 10
0021-9193/00/$04.00+0
Copyright © 2000, American Society for Microbiology. All rights reserved.
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